Expression and screening in yeast of genes mutagenized in vitro.

摘要: Publisher Summary This chapter describes two methods that allow the expression of mutant alleles in yeast. One method produces mutations reside chromosome and other on plasmids. Yeast plasmids are grouped into several classes based their mode replication. All commonly used yeast shuttle vectors; they can be propagated both Escherichia coli. integrating (YIp) contain a gene as selectable marker for transformation. These introduced virtually any genetically marked strain. Chromosomal genes exhibit low reversion frequencies, allowing wild-type copies these to markers The procedure generating chromosomal depends properties YIp plasmids, which do not sequences autonomous replication only following integration chromosome. They integrate by recombination between carried plasmid homologous present genome.